Methods for treatment of benign and malignant tumors, including lymphomas, leukemias, and leiomyomas

ABSTRACT

In a preferred embodiment, drugs having chemotherapeutic properties which are useful against certain neoplastic disorders with wide safety margins as evidenced by their low toxicity, and molecular actions. Such drugs include as active ingredient(s) one or more N-substituted 2-(1H) pyridone(s) and/or N-substituted 3-(1H) pyridone(s). The compositions of this invention are novel as anti-neoplastic drugs, namely as an agent for treating leukemias, lymphomas, and leiomyomas  as agent( s )  for treating benign and malignant tumors, including lymphomas, leukemias, and leiomyomas.

This application is a CIP of Ser. No. 09/162,011 filed Sep. 28, 1998which is a CIP of Ser. No. 08/913,202 filed Sep. 3, 1997 ABN., which isa CIP of PCT/US96/02737 filed Mar. 4, 1996 which is a CIP of Ser. No.08/397,962 filed Mar. 3, 1995 ABN.

BACKGROUND OF THE INVENTION

1. Field of the Invention

The present invention relates to medical compositions and methods forthe chemotherapeutic treatment of lymphomas, leukemias, and leiomyomasgenerally and, more particularly, to compositions comprising one or moreN-substituted 2(1H) pyridones and/or one or more N-substituted 3(1H)pyridones as active ingredient(s). The selected compounds nay be usedalone or as an adjunct to other forms of neoplastic therapy includingsurgery, other chemotherapeutic compounds, radiation therapy, andimmunotherapeutic agents.

2. Background Art

The causes of leukemias and leiomyomas are poorly understood, arecomplex, and involve interplay between the basic genetic material in thenucleus of cells. An abnormal reaction of cellular genetic DNA tointernal or external factors can create a new deviation in the cellgenetic code, or in the genetic DNA generated communication proteinswhich creates neoplastic perturbations in the transcription processgoverning the specific cell cycle stages of otherwise normal celldivision, and proliferation.

Cell proliferation is defined as the increase in number of cellsresulting from completion of the cell cycle, as contrast to growth,which is the increase in the individual cell mass.

Extracellular or intracellular factors can determine whether a quiescentcell will begin to proliferate and also whether a normal proliferatingcell in phase G1 will begin to cycle or will revert to quiescence. Aftercells enter into the S Phase, cell-cycle events become largelyindependent of prior extracellular factors., while they go on to divideand produce two daughter cells.

Among the carcinogenic factors of external origin, acting internally,are physical carcinogens such as ionizing or ultraviolet radiation, andthe presence of foreign substances such as asbestos. Carcinogenicsubstances acting internally include various chemicals, natural orman-made, which can effect directly or indirectly cell DNA to elicitintracellular oncogenic events. In addition biological substances suchas bacteria, viruses, parasites, hormones and cytokines have beenimplicated in mammalian carcinogenesis.

In benign and malignant tumors including lymphomas, leukemias andleiomyomas, the control of proliferation is deranged. After induction ofaltered proliferation control, deranged cell differentiation isinitiated in phase G1, and is a hallmark of neoplastic cells (Pardee. A.B., Science, Nov. 3, 1989, p.603).

Neoplasms are manifest when the normal progression of the orderlyrelationship between cell division and cell differentiationmalfunctions. With the usual cell division sequence in normal cells, theproliferation of cells is restricted to non-differentiated stem cellswhich ordinarily differentiate and reproduce to provide a replacementfor aged dying cells.

Neoplasms arising from lympho-hematopoietic origin may be identified asleukemias, lymphomas, leiomyomas, etc.

The term “anti-neoplastic” or “anti-tumor” refers herein to the (a)chemotherapeutic inhibition of arrest of the growth, and (b) thedestruction of mammalian benign , (for example leiomyomas) and/ormalignant tissues (such as leukemias, or lymphomas), and leiomyomasfound in various organs and tissues of the body.

Although most tissues and organs of the human body may becomeneoplastic, the basic processes leading to diverse tumors appear to bequite similar. Normal cells proliferate or reproduce in rigorouscompliance with programmed guidance from parental or adjacent cells.Such unceasing, disciplined instruction ensures that each tissuemaintains a size, architecture and function appropriate to the body'sneeds.

Neoplastic cells, in distinct contrast, become unresponsive to the usualcontrols of parental or adjacent cells with respect to proliferation,architecture and/or function. These neoplastic cells frequently (a)migrate from the site where they began, (b) invade nearby tissues, and(c) travel through the blood and lymphatic circulatory systems to formneoplastic lesions at distant sites in the body. These lesions becomelethal when they disrupt the normal function of other tissues or organsessential for the patient survival.

Multiple genetic changes occur during the transformation of normal cellsinto neoplastic cells. This is facilitated in neoplastic cells by lossof fidelity in the processes that replicate, repair, and segregate thegenome structure. Advances in our understanding of the cell cycle revealhow fidelity is normally achieved by the coordinated activity ofcyclin-dependent kinases, checkpoint controls, and repair pathways, andhow this fidelity can be abrogated by specific genetic changes. Therecognition of molecular mechanisms for cellular transformation may helpidentify the mechanisms by which chemotherapeutic compounds are usefulin the treatment of neoplastic diseases (Hartwell, L. and Kasten, M.,1994, Science. 266:1821-1828).

Control systems enforcing interdependency in the cell cycle are called“checkpoints.” Elimination of checkpoints can result in cell death,infidelity in the distribution chromosomes or other cellular organelles,or increased susceptibility to external perturbations such as DNAdamaging agents. Such perturbations can result in neoplastictransformation of cells and tissues (Hartwell, L. and Weinert, T., 1989,Science, 246:629-634).

The cell-type-specific expression of most genes is determined at thetranscription level. Transcription factors are involved in the controlof the process. To understand the basis of this regulation, it hasbecome important to analyze the control of transcription factorsthemselves. A variety of transcriptional, translational, andpost-translational mechanisms have been described. The most direct wayfor a cell to regulate the abundance of a factor is to adjust theproduction of the mRNA encoding it. Thus far, the control of manycell-type and tissue-specific transcription factors has been found tooccur at the transcription level (Falvey, E., and Schibler, U., FASEBJ., 1991, 5:309-314).

Recent studies indicate that extracellular signals often effect cellproliferation and differentiation by modulating intracellulartranscription factor activity via protein phosphorylation cascades,which involves the transduction systems used to transmit information(signals) from the cell surface to the transcription machinery of thecell nucleus (Karin, M., 1992, FASEB J. 6:2581-2590).

Cell nuclear transcription factors regulate tissue and stimulus-specificgene expression through their ability to integrate extracellular signalsat the nucleus. Several human diseases, including neoplasias,cardiovascular disease, and neurological and autoimmune disorders,result from aberrations in the expression of genes regulated by thesetranscription factors (Manning, A. Gonzales, R., and Bennett, B., ExpertOpin. Ther. Pat., 1997, 7:225-231).

Normally, the body's tissues prevent excessive proliferation of cells bydepriving them of excessive amounts of growth-stimulating factors, or byflooding the cells with antiproliferative factors derived from adjacentor parental cells which block the actions of the growth stimulatingfactors.

Certain cellular proteins, through their intrinsic ability to regulate ahost of other genes involved in the control of cell proliferation, canreorganize and redirect a cell's normal or abnormal fate. Thus, the lossof these growth controlling genes by deletion or mutation is a commonoccurrence in neoplasias (Lozano, G. and Hulboy, D. L., Methods (SanDiego) 1995, 8:215-224.)

Some cell cycle derangements stem from extracellular influences. Manyneoplasia causing oncogenes, for example, turn out to encode componentsof the pathways through which various growth factor signals feed intothe cell cycle to stimulate cell division. This is an importantdemonstration that the protein encoded by the p53 tumor suppresser geneinhibits cell growth by turning on the production of a specializedprotein that blocks the cell cycle. The intracellular gene encoding onecomponent of the cell cycle machinery, a protein called cyclin D1, aswell as several others, are oncogene candidates. A significant amount ofexperimental evidence indicates that excess cyclin D1 causes neoplasias,but additional data suggest that several gene changes are implicated inthe transformation of normal cells into a neoplastic configuration. Forexample, a hallmark of normal healthy cells is their ability todifferentiate, but neoplastic cells cannot differentiate due theblockade of this response via the combined action of cyclins D2 and D3(Marx, J., 1994, Science, 263:319-321).

Other recent data strongly suggest that deregulation of the restrictioncheckpoints in the cell-cycle G-1 phase is required for the transitionto neoplastic disarray as seen in neoplasias (Strauss, M.; Lucas, J.;Bartek, J. Nat. Med. (N.Y.), 1995 1:1245-1246).

Mitogenic stimulation of normal cells initiates a sequence of eventsleading to activation of cyclin-dependent kinases, phosphorylation ofRb, and subsequent entry into of the cell into the S phase. Many typesof neoplasms have lost sensitivity to the growth-inhibitory actions ofTGF-beta-1, and this may derive from dysregulated expression of cyclin,cdk, and cdk inhibitor genes (Satterwhite, D. and Moses, H., 1994-1995(Publ. 1995), Invasion Metastasis, 14:309-318).

Large T-antigen expression in human fibroblasts selectively uncouplescyclin D1 from cdk4, and subsequent immortalization of these cellsresults in additional changes in the cyclin D-dependent cell cycleregulatory pathways (Peterson, S. et al., 1995, Cancer Res.,55:4651-4657).

Representative Neoplasms of the Immune System

Each neoplasm of the immune system exhibits a distinct clinical andpathologic character, yet these disorders share a number of commonfeatures. Systemic symptoms of fever, night sweats, and weight loss maybe present and are associated with advanced stages of the disease. Thesetumors usually appear in one or more organs of the hematopoietic system(lymph nodes, spleen, liver, bone marrow). If untreated, fataldissemination to all of these organs, as well as other sites occurs.

Chronic myeloid leukemia (CML) usually presents clinically in a chronicphase of variable duration, after which a fatal condition similar toacute leukemia (blast crisis) develops. The problem of initial therapyfor chronic phase CML is that current conventional therapy offers littlechance of long-term survival of the patient with no chance of cure,whereas an agent that offers superior survival and a chance of cure isvery toxic and expensive (Kattan, M., et al., Ann. Intern. Med., 1996;125:541-548).

The disease is characterized by overproduction of granulocytic cells(especially neutrophilic types), leading to marked splenomegaly, andvery high white blood cell counts. Rises in basophils and thrombocytosisoccur frequently.

Inhibition of proliferation by pirfenidone of human uterine leiomyomasor fibroid cells in vitro. According to the literature, leiomyomas arethe most common pelvic tumors in women with a reported incidence of 20%or more (Merrill, J., Creasman, W. 1990, “Danforth's Obstetrics andGynecology,” Scott, J., et al., eds., 6th edition. Philadelphia.Lippincott, pp. 1023-1039). Most common symptoms associated with thesebenign tumors are excessive abnormal uterine bleeding, pelvic pain,infertility and increased urinary frequency. Consequently, the presenceof leiomyomas is the leading cause for hysterectomy in the United States(Wilcox, L. et al., 1994, Hystersectomy in the United States, 1990,Obstet. Gynecol. 83:549-555).

The majority of chemotherapeutic neoplasia agents in current clinicalpractice are toxic compounds and exert their greatest anti-neoplasiaeffect when employed at the maximum tolerated dose. With thesechemotherapeutic agents, toxic actions to normal tissue can greatlylimit the amount that can be safely administered. To date, the mostcommonly utilized agents are only partially selective in their toxicity.Thus, they are damaging to both normal and neoplastic cells. Theseagents disrupt major intracellular systems such as DNA synthesis andessential enzymes systems. Nevertheless treatment of neoplastic diseaseis predicated on exploiting the small differences between healthy normalcells and neoplastic cells.

Accordingly, it is a principal object of the present invention toprovide compositions for the inhibition or arrest of the growth or forthe destruction of mammalian benign (for example leiomyomas) and/ormalignant tumors lymphomas, (for example leiomyomas, and leukemias,etc.).

It is a further object of the present invention to provide suchcompositions that provide a means of (1) arresting the proliferation ofand (2) then killing the abnormal cells of neoplastic tissue withoutserious or fatal injury to healthy normal cells and tissues.

It is an additional object of the invention to provide such compositionsthat comprise one or more N-substituted 1-(1H) pyridone(s) and/orN-substituted 3-(1H) pyridone(s) as active anti-tumor ingredient(s).

Other objects of the present invention, as well as particular featuresand advantages thereof, will be elucidated in, or be apparent from, thefollowing descriptions.

SUMMARY OF THE INVENTION

The present invention achieves the above objects, among others, andovercomes the limitations of the prior art by providing, an in apreferred embodiment, drugs having chemotherapeutic properties which areuseful against certain neoplastic disorders with wide safety margins asevidenced by their low toxicity, and molecular actions. Such drugsinclude as active ingredient(s) one or more N-substituted 2-(1H)pyridone(s) and/or N-substituted 3-(1H) pyridone(s). The compositions ofthis invention are novel as anti-neoplastic drugs, namely as an agentfor treating malignant neoplasms (such as leukemias , and lymphomas,and/or benign neoplasms (such as leiomyomas).

BRIEF DESCRIPTION OF THE DRAWING

FIG. 1 is a bar graph showing the inhibition by pirfenidone of theproliferation of human pro-myelocytic leukemia cells.

FIG. 2 is a bar graph showing the inhibition by prifenidone of humanT-cell leukemia cells.

FIG. 3 is a bar graph showing the inhibition by pirfenidone ofproliferation of human Burkitt's lymphoma cells.

FIG. 4 is a bar graph showing the inhibition by pirfenidone of humanleiomyoma cells.

DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS Methods for EvaluatingAnti-Cancer Agents in the Present Invention

All-trans-retinoic acid is known to specifically induce fresh humanpromyelocytic leukemia cells (AML3) to differentiate in vitro to maturefunctional gramulocytes which loose their self-renewal potency andspontaneously die. These results were confirmed in vivo: AML-3 patientstreated with oral all-trans retinoic acid alone achieve completeremission (Chomienne, Ch., “Retinoids: Basic Sci. Clin. Appl.,” 1994,p.233-241. Edited by Livrea and Vidali, Birkhaeuser, Basel,Switzerland).

Evidence that in vitro prednisolone resistance is of prognostic value inchildhood acute lymphoblastic leukemia (ALL) was explored further inadult ALL leukemia. Blast cells from 30 patients were exposed toprednisolone (PDN) (0.1) microMol to 35 microMol), and cytotoxicity wasassayed calorimetrically with soluble tetrazolium formazan (2,3-bis(2-methoxy-4-nitro-5-sulfophenyl)-5-[(phenylamino)carbamyl]-2H-tetrazoliumhydroxide, XTT). The IC50 varied greatly among the samples from 0.3microMol to >35 microMol. 15 microMol was subsequently chosen as IC50cutoff between in vitro resistant and sensitive cases. By regressionanalysis, PDN-induced cytotoxicity was found to be significantly relatedto apoptosis. However, correlation with clinical symptoms was notdemonstrated. Nevertheless, disease free survival was significantlybetter in sensitive patients (Tosi, P. et al., Eur. J. Haematology,1996, 52:134-141.)

Chronic lymphocytic leukemia (CLL) is characterized by delayedsenescence and a slow accumulation of small lymphocytes. Elevatedcytokine levels have been detected in urine from patients with a varietyof neoplastic diseases including various leukemias. The source of theelevated levels of cytokine has not been determined. In the presentstudy, the intracellular cytokine level in lymphocytes from 36 patientswith B-CLL and 15 normal donors was determined using an enzyme-linedimmunoassay. In cells derived from patients with high risk disease, themedian level of cytokine was 382 pg/200,000 cells compared to 91pg/200,000 cells in patients with intermediate disease. In patients withlow-risk disease, median cytokine level was 4.9 pg/200,000 cells, and innormal controls, it was 6.0 pg/200,000 cells. The respective differenceswere statistically significant (Menzel et al., Blood, 1996,87:1056-1063).

In 1996, approximately 3,500 new cases (mostly children) of acutelymphoblastic leukemia (ALL) were diagnosed in the US. Currentchemotherapy induces complete remission in many patients, but 20% suffera critical relapse.

It has been discovered by the present inventor that pirfenidone andother N-substituted 2(1-H) pyridone compounds and N-substituted 3(1H)pyridone compounds have anti-tumor activity. Heretofore, before thediscovery of the invention disclosed herein, no effective N-substitutedpyridone agent or composition has been available for the inhibition andarrest of the growth, or for the destruction of mammalian benign ormalignant tissues such as found in leukemias, lymphomas or leiomyomas.

The “anti-tumor” activity discovered by the present inventor and as usedherein refers to the ability of an active substance to inhibit or arrestthe life-threatening proliferation of neoplastic cells in benign andmalignant tumors including lymphatic or hymelogenous leukemias,lymphomas, and leiomyomas.

Methods of preparation of some N-substituted 2(1-H) pyridones useful inthe present invention are described in U.S. Pat. No. 3,839,346, issuedOct. 1, 1974, to Gadekar, and titled N-SUBSTITUTED PYRIDONE AND GENERALMETHOD FOR PREPARING PYRIDONES, the disclosure of which is incorporatedby reference hereto.

EXAMPLE I

Inhibition of proliferation by pirfenidone of human promyelocyticleukemia (HL-60) cells in vitro.

Pirfenidone was suspended in RPMI, 2.0% FBS, at 100 micrograms to 900micrograms per ml, and one other culture served as a control. Five ml ofeach pirfenidone concentration, and of the control were placed into aT25 tissue culture flask along with 1×10⁵ cells. Every day for 4 days analiquote from each flask was taken out, then stained with Trypan Blue,and counts were repeated three times on each aliquote.

The inhibition of proliferation of HL-60 cells by pirfenidone wasdirectly related to the graded concentrations of pirfenidone in therespective culture flasks. After 3 and 4 days at a concentration of 900micrograms/ml, a 63% pharmacologic inhibition was maintained withoutcausing a toxic death of the cells (see FIG. 1). Inhibition ofproliferation as well as deaths of almost all cells occur withpirfenidone concentrations at 1400 micrograms/ml.

EXAMPLE II

Inhibition of proliferation by pirfenidone of human T-cell leukemia(JURKAT) cells in vitro.

To determine the inhibitory effect of pirfenidone concentrations onhuman T-cell leukemia (JURKAT), a MTS assay was used which incorporatesa tetrazolium dye that measures the cellular conversion of the dye intoa formazan product by the activation of NADH-generating dehydrogenasesfound in metabolically active cells.

Procedure

1. 100 microl. of cells was seeded into 96 well plates at 1×10⁵ cells/mlin media containing 100 mcg/ml, 300 mcg/ml and 900 mcg/ml ofpirfenidone, as well as a pirfenidone-free media control.

2. Cells were incubated at 37 degrees C., at 5.0% CO2 for 72 hours.

3. After incubation a standard curve was made of 100 microliters ofcells added to the plate at 5×10⁵ cells/ml making 1:2 dilutions all theway down to a pre-set degree of dilution.

4. 2.0 mls of the MTS solution was mixed with a 100 microliter of thePMS solution, and 20 microliters of this combined dye solution was addedto each well and incubated at 37 degrees C., 5.0% CO2 for 3 hours.

5. Subsequently the absorbance was read at 490 nm.

Exposure for only 72 hours of human T-cell leukemia (JURKAT) cells, at900 mcg/ml of pirfenidone caused a statistically significant 75%reduction in number of cell which were active (75% no longer were ableto proliferate; FIG. 2).

EXAMPLE III

Inhibition of proliferation by pirfenidone of human Burkitt's lymphoma(RAJI) cells in vitro.

To determine the inhibitory effect of pirfenidone concentrations onBurkitt's lymphoma (RAJI) cells, a MTS assay was used which incorporatesa tetrazolium dye that measures the cellular conversion of the dye intoa formazan product by the activation of NADH-generating dehydrogenasesfound in metabolically active cells.

Procedure

1. 100 microl. of cells was seeded into 96 well plates at 1×10⁵ cells/mlin media containing 100 mcg/ml, 300 mcg/ml and 900 mcg/ml ofpirfenidone, as well as a pirfenidone-free media control.

2. Cells were incubated at 37 degrees C., at 5.0% CO2 for 72 hours.

3. After incubation a standard curve was made of 100 microliters ofcells added to the plate at 5×10⁵ cells/ml making 1:2 dilutions all theway down to a pre-set degree of dilution.

4. 2.0 mls of the MTS solution was mixed with a 100 microliter of thePMS solution, and 20 microliters of this combined dye solution was addedto each well and incubated at 37 degrees C., 5.0% CO2 for 3 hours.

5. Subsequently the absorbance was read at 490 nm.

Exposure for only 72 hours of these human Burkitt's lymphoma (RAJI)cells, at 900 mcg/ml of pirfenidone caused a statistically significant45% reduction in number of cells which were active (45% no longer wereable to proliferate; FIG. 3).

EXAMPLE IV

Leiomyoma tissues were obtained from several premenopausal women withsymptomatic uterine fibroids at elective hysterectomy, and who were notreceiving any hormonal or other drug therapy. The fibroid tissues wereminced into 1-2 mm# explants and placed in suitable DMEM supplementedwith 10% bovine serum. The fibroid tissue was digested for 14-18 hoursat 37 degrees C. in an incubator. After centrifugation, a resulting cellpellet was resuspended in DMEM and then placed in culture flasks. Thecultures were maintained at 37 degrees centigrade in a humidifiedatmosphere of 5% CO2 and 95% air.

For determining cell proliferation utilizing tritiated thymidineincorporation as an assay, leiomyoma cells were cultured in 96-wellplates (15,000 cells/well) for 48 hours in DMEM plus 10.0% FBS serum.Cells were then made quiescent by culturing in DMEM plus 0.5% FBS serumfor 48 hours. These quiescent cells were washed, and then placed in DMEMplus 10.0% FBS serum containing graded concentrations of pirfenidone(10.0, 100, 300 and 1000 micrograms/ml). After 18 hours, the cellsreceived 0.2 uCi/well of [3H]-thymidine, and the incubation wascontinued for 6 more hours. Subsequently, cells were harvested andcounted in a beta-counter to the rate of incorporation of[3H]-thymidine. Results are summarized in Table 1.

TABLE 1 INHIBITION BY PIRFENIDONE OF PROLIFERATION OF HUMAN LEIOMYOMACELLS PIRFENIDONE RECENT DEAD TREATMENT CELL COUNTS (DAY 7) CELLS (DAY7) (CONC/ML) LEIOMYOMA LEIOMYOMA 0.0 MG 380,000 9.0 0.01 MG 315,000 8.00.1 MG 205,000 10.0 0.3 MG 170,000 6.0 1.0 MG 69,000 16.0

EXAMPLE V

Leiomyoma cells were plated in 100 mm dishes (100,000 each dish) andallowed to attach overnight in DMEM plus 10% fetal bovine serum untilthey reached 80-90% confluence. The following day all cells receivedDMEM plus 10% fbs containing various concentrations of pirfenidone (0.0micrograms, 100 micrograms, 300 micrograms or 1000 microgram/ml forseven (7) days. The medium was changed with addition of fresh treatmentson days 3 and 5. On day 7, cells were harvested and counted. Cellviability was assessed using the Trypan Blue exclusive stain. Resultsare displayed in FIG. 4.

A significant inhibitory effect on proliferation was seen for leiomyomacells exposed to graded concentrations of pirfenidone in the culturemedia (FIG. 4). A significant increase in the percentage (16%) of deadleiomyoma cells was found at the 1000 micrograms/ml concentration ofpirfenidone (Table 1).

The data demonstrates the anti-tumor effect of on leiomyomas indicateindicates that pirfenidone will be useful in eliminating the most severesymptoms associated with leiomyoma tumors (excessive abnormal arterialbleeding, pelvic pain, infertility and increased urinating frequency).

These compounds also can be employed in combination to enhance othertypes of therapy (surgery, radiation, immunotherapy or otherchemotherapeutic compounds). Another facet of the present invention isabsence of any severely debilitating adverse effects. The absence ofsuch severe toxic reactions reduces or eliminates the patient discomfortinherent in conventional treatments for neoplasias. The delivery methodto patients being treated may consist of oral, intramuscular orintravenous administration.

The N-substituted pyridones of the present invention pharmacologicallyarrest the proliferation of leukemias, Llymphomas, or leiomyoma cells ortissue, at concentrations that are ⅓rd- 1/10th that which is toxic orkilling to these neoplastic cells. Accordingly, this inventioncharacterizes a group of chemotherapeutic compounds which provide ameans of (1) arresting the proliferation and (2) then destroying themalignant cells by raising the pirfenidone concentration of the cell oftissue to the cytotoxic level or by adding another known cytotoxicanti-neoplastic agent, the neoplastic cells can be destroyed oreradicated without serious or fatal injury to healthy normal cells andtissues.

These compounds also can be employed in combination to enhance othertypes of therapy (surgery, radiation, immunotherapy or otherchemotherapeutic compounds).

Another facet of the present invention is absence of any severelydebilitating generalized adverse effects. The absence of such severetoxic reactions reduces or eliminates the patient discomfort inherent inconventional neoplasia treatments.

The differences in the concentrations of pirfenidone whichpharmacologically arrest the proliferation of neoplasia neoplastic cellsand the pirfenidone concentration which kills neoplasia cells affords alarger margin of safety for patients, (Raghu, G., et al., Amer. J. Resp.and Critical Care Med., 1997, Vol. 155:A741), and thereby distinctlyreduces the incidence of serious adverse effects experienced by patientsduring treatment, as compared to treatment with currently conventionalanti-neoplasia anti-neoplastic agents.

The intracellular action of pirfenidone in (1) arresting theproliferation and (2) subsequent destruction of the abnormal orneoplastic cells takes place in the cell nucleus and directly involvesthe signaling via the specific gene activated proteins (for example,p53, Rb, WT1, etc.) and ameliorating or blocking the impact of such geneproteins on the cell transcription apparatus and cyclins. These specificgene proteins act on the check points of the cell cycle to prevent orcorrect the aberrant gene protein signals impacting on the cyclins andcheck points.

It is estimated that the effective human dosage of one or moreN-substituted 2-(1H) pyridone(s) and/or N-substituted 3-(1H) pyridone(s)in practicing the present invention is from about 250 to about 750 mg/kgof body weight per day, which dosage may be taken in the diet.

The general structural formula of N-substituted 2-(1H) pyridones is:

where: R1 is selected from the group consisting of (1) an alkyl group,with R3 hydrogen, and (2) hydrogen, with R3 consisting of an alkylgroup; A is an aryl group; and R2 and R4 are hydrogen.

The general structural formula for the N-substituted 3-1(H) pyridonesis:

where: R2 is selected from the group consisting of (1) an alkyl group,with R3 hydrogen, and (2) hydrogen, with R3 consisting of an alkylgroup; A is an aryl group; and R1 and R4 are hydrogen.

Examples of the pyridone compounds which have been found or are believedto be effective in practicing the present invention include:

-   -   5-Methyl-1-phenyl-2-(1H) pyridone    -   5-Methyl-1-(3-nitrophenyl-2)-(1H) pyridone    -   5-Methyl-1-(4′-methoxyphenyl)-2-(1H) pyridone    -   5-Methyl-1-p-tolyl-2-(1H) pyridone    -   5-Methyl-1-(3′-trifluoromethylphenyl)-2-(1H) pyridone    -   1-(4′Chlorophenyl)-5-methyl-2-(1H) pyridone    -   5-Methyl-1-(2′-naphthyl)-2-(1H) pyridone    -   5-Methyl-1-(′-naphthyl)-2-(1H) pyridone    -   3-Methyl-1-phenyl-2-(1H) pyridone    -   6-Methyl-1-phenyl-2-(1H) pyridone    -   3,6-Dimethyl-1-phenyl-2-(1H) pyridone    -   5-Methyl-1-(2′thienyl)-2-(1H) pyridone    -   1-(2′-Furyl)-5-methyl-2-(1H) pyridone    -   5-Methyl-1-(5′-quinolyl)-2-(1H) pyridone    -   5-Methyl-1-(4′-pyridyl)-2-(1H) pyridone    -   5-Methyl-1-(3′-pyridyl)-2-(1H) pyridone    -   5-Methyl-1-(2′-pyridyl)-2-(1H) pyridone    -   5-Methyl-1-(2′-quinolyl)-2-(1H) pyridone    -   5-Methyl-1-(4′-quinolyl)-2-(1H) pyridone    -   5-Methyl-1-(2′-thiazolyl)-2-(1H) pyridone    -   1-(2′-Imidazolyl)-5-methyi-2-(1H) pyridone    -   5-Ethyl-1-phenyl-2-(1H) pyridone    -   3-Ethyl-1-phenyl-2-(1H) pyridone    -   1-Phenyl-2-(1H) pyridone    -   1-(4′-Nitrophenyl)-2(1H) pyridone    -   5-Methyl-3-phenyl-1-(2′-thienyl)-2-(1H) pyridone    -   5-Methyl-1-phenyl-3-(1H) pyridone    -   5-Methyl-1-(4′-methoxyphenyl)-3-(1H) pyridone    -   5-Methyl-1-p-tolyl-3-(1H) pyridone    -   1-(4′-Chlorophenyl)-5-methyl-3-(1H) pyridone    -   5-Methyl-1-(2′-naphthyl)-3-(1H) pyridone    -   4-Methyl-1-phenyl-3-(1H) pyridone    -   6-Methyl-1-pheyl-3-(1H) pyridone    -   5-Methyl-1-(2′-thienyl)-3-(1H) pyridone    -   1-(2′-Furyl)-5-methyl-3-(1H) pyridone    -   5-Methyl-1-(5′-quinolyl)-3-(1H) pyridone    -   5-Methyl-1-(3′-pyridyl)-3-(1H) pyridone    -   5-Methyl-1-(2′-pyridyl)-3-(1H) pyridone    -   5-Methyl-1-(2′-quinolyl)-3-(1H) pyridone    -   5-Ethyl-1-phenyl-3-(1H) pyridone    -   1-Phenyl-3-(1H) pyridone.

It will thus be seen that the objects set forth above, amoung thoseelucidated in, or made apparent from, the preceding description, areefficiently attained and, since certain changes may be made in the abovecompositions and methods without departing from the scope of theinvention, it is intended that all matter contained in the foregoingdisclosure shall be interpreted as illustrative only and not in alimiting sense.

It is also to be understood that the following claims are intended tocover all of the generic and specific features of the invention hereindescribed and all statements of the scope of the invention which, as amatter of language, might be said to fall therebetween.

1. A method of treating lymphomas, leukemias, and/or leiomyomas one or more malignant tumors and/or one or more benign tumors in a laboratory animal or a human, comprising: administering to said laboratory animal or said human an effective dose of a composition including one or more pharmaceutical substances selected from the group consisting of N-substituted 2-(1H) pyridones, N-substituted 3-(1) pyridones, and pharmaceutically acceptable salts thereof, wherein said 2-(1H) pyridones have the following general structural formula:

where: R1 is selected from the group consisting of (1) an alkyl group, with R3 hydrogen, and (2) hydrogen, with R3 consisting of an alkyl group; A is an aryl group; and R2 and R4 are hydrogen; and wherein said 3-(1H) pyridones have the following general structural formula:

where: R2 is selected from the group consisting of (1) an alkyl group, with R3 hydrogen, and (2) hydrogen, with R3 consisting of an alkyl group; A is an aryl group; and R1 and R4 are hydrogen.
 2. A method, as defined in claim 1, wherein: said composition is administered orally or parenterally to said laboratory animal at a rate of from about 250 to about 750 mg/kg of body weight per day.
 3. A method, as defined in claim 1, wherein: said composition is administered orally or parenterally to a said human at a rate of from about 20 to about 60 mg/kg of body weight per day.
 4. A method of treating lymphomas, leukemias, and/or leiomyomas one or more malignant tumors and/or one or more benign tumors in a laboratory animal or a human, comprising: administering to said laboratory animal or said human an effective dose of a composition including one or more pharmaceutical substances selected from the group consisting of N-substituted 2-(1H) pyridones, N-substituted 3-(1H) pyridones, and pharmaceutically acceptable salts thereof, said N-substituted 2-(1H) pyridones and said N-substituted 3-(1H) pyridones being selected from the group consisting of: 5-Methyl-1-phenyl-2-(1H) pyridone, 5-Methyl-1-(3-nitrophenyl-2)-(1H) pyridone, 5-Methyl-1-(4′-methoxyphenyl)-2-(1H) pyridone, 5-Methyl-1-p-tolyl-2-(1H) pyridone, 5-Methyl-1-(3′-trifluoromethylphenyl)-2-(1H) pyridone, 1-(4′Chlorophenyl)-5-methyl-2-(1H) pyridone, 5-Methyl-1-(2′-naphthyl)-2-(1H) pyridone, 5-Methyl-1-(1′-naphthyl)-2-(1H) pyridone, 3-Methyl-1-phenyl-2-(1H) pyridone, 6-Methyl-1-phenyl-2-(1H) pyridone, 3,6-Dimethyl-1-phenyl-2-(1H) pyridone, 5-Methyl-1-(2′thienyl)-2-(1H) pyridone, 1-(2′-Furyl)-5-methyl-2-(1H) pyridone, 5-Methyl-1-(5′-quinolyl)-2-(1H) pyridone, 5-Methyl-1-(4′-pyridyl)-2-(1H) pyridone, 5-Methyl-1-(3′-pyridyl)-2-(1H) pyridone, 5-Methyl-1-(2′-pyridyl)-2-(1H) pyridone, 5-Methyl-1-(2′-quinolyl)-2-(1H) pyridone, 5-Methyl-1-(4′-quinolyl)-2-(1H) pyridone, 5-Methyl-1-(2′-thiazolyl)-2-(1H) pyridone, 1-(2′-Imidazolyl-5-methyl-2-(1H) pyridone, 5-Ethyl-1-phenyl-2-(1H) pyridone, 3-Ethyl-1-phenyl-2-(1H) pyridone, 1-Phenyl-2-(1H) pyridone, 1-(4′-Nitrophenyl)-2-(1H) pyridone, 5-Methyl-3-phenyl-1-(2′-thienyl)-2-(1H) pyridone, 5-Methyl-1-phenyl-3-(1H) pyridone, 5-Methyl-1-(4′-methoxyphenyl)-3-(1H) pyridone, 5-Methyl-1-p-tolyl-3-(1H) pyridone, 1-(4′-Chlorophenyl)-5-methyl-3-(1H) pyridone, 5-Methyl-1-(2′-naphthyl)-3-(1H) pyridone, 4-Methyl-1-phenyl-3-(1H) pyridone, 6-Methyl-1-phenyl-3-(1H) pyridone, 5-Methyl-1-(2′-thienyl)-3-(1H) pyridone, 1-(2′-Furyl)-5-methyl-3-(1H) pyridone, 5-Methyl-1-(5′-quinolyl)-3-(1H) pyridone, 5-Methyl-1-(3′-pyridyl)-3-(1H) pyridone, 5-Methyl-1-(2′-pyridyl)-3-(1H) pyridone, 5-Methyl-1-(2′-quinolyl)-3-(1H) pyridone, 5-Ethyl-1-phenyl-3-(1H) pyridone, and 1-Phenyl-3-(1H) pyridone. 